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白血病抑制因子 (LIF)是一种多功能细胞因子 ,在生物发育及维持正常生理功能中发挥着重要的作用。以成人血细胞的总RNA为模板 ,利用RT PCR方法从成人外周血细胞中扩增人白血病抑制因子 ,而后将其克隆到真核表达载体pcDNA3,在哺乳动物细胞中成功表达。将分泌到细胞培养基中的LIF因子收集 ,通过LIF信号转导通路下游蛋白质STAT3磷酸化水平的检测、EMSA实验以及荧光酶报告系统检测 ,发现其具有激活STAT3信号通路的生物学活性。同时 ,[3H] TdR参入实验的结果也表明 ,所表达LIF因子能显著抑制鼠骨髓白血病细胞系M1的增殖。
Leukemia inhibitory factor (LIF) is a multifunctional cytokine that plays an important role in the biological development and maintenance of normal physiological functions. Using the total RNA of adult blood cells as a template, human leukemia inhibitory factor was amplified from adult peripheral blood cells by RT PCR and cloned into the eukaryotic expression vector pcDNA3 for successful expression in mammalian cells. The LIF secreted into the cell culture medium was collected. The phosphorylation level of STAT3 protein downstream of the LIF signal transduction pathway was detected by EMSA assay and luciferase reporter system. The LIF factor secreted into the cell culture medium was found to activate the biological activity of STAT3 signaling pathway. At the same time, the results of the incorporation of [3H] TdR also showed that the expressed LIF factor significantly inhibited the proliferation of murine myeloid leukemia cell line M1.